Method and composition for treating urinary tract infections

ABSTRACT

A method for the treatment of urinary tract infections and a composition for use in such method, such method comprising administering to a patient suffering from a urinary tract infection an effective anti-bacterial dosage of a composition comprising in combination: 
     A. a source of methenamine, e.g. methenamine; and 
     B. a source of hydroxamate groups, e.g., acetohydroxamic acid. 
     The source of hydroxamate groups is administered in the amount effective to potentiate the anti-bacterial effect of the source of methenamine. The method is specifically applied to the treatment of urinary infections caused by urease producing bacteria, especially bacteria of the species proteus. 
     In addition, where colonization of the bacteria can be tolerated and where it is only necessary to eliminate the pathogenicity of the urease producing bacteria induced infection, effective treatment can be achieved by administration of only the source of hydroxamate groups, preferably acetohydroxamic acid. Also, by maintaining the urine at an acid pH in the presence of urease producing bacteria by administering a source of hydroxamate groups, struvite and apatite urinary stones normally associated with urease producing bacteria can be dissolved.

RELATED APPLICATION

This application is a continuation-in-part of application Ser. No.441,245, filed Feb. 11, 1974, now abandoned.

BACKGROUND OF THE INVENTION

1. Field of Invention

This invention relates to a method for treating urinary tract infectionsand a composition for use in such method; and more specifically thepresent invention relates to such method and composition for thetreatment of urinary tract infections, specifically urinary tractinfections caused by bacteria that produce urease, especially those ofthe species Proteus, wherein the anti-bacterial effect of methenamine ispotentiated or synergized by administering a source of methenamine incombination with a source of hydroxamate groups. In addition, thisinvention relates to a method of eliminating the pathogenicity of ureaseproducing bacteria by administering to a patient suffering from aninfection caused by urease producing bacteria a source of hydroxamategroups. Furthermore, the present invention provides a method ofdissolving struvite and apatite urinary stones generally associated withurease producing bacteria by administering a source of hydroxamategroups which provides urine of physiological pH, which due tounder-saturation with respect to struvite and apatite crystals caneffectively dissolve the associated stones.

2. Description of the Prior Art

The increasing awareness of the importance of urinary tract infectionshas brought about the realization that adequate drug therapy is requiredfor the treatment of the infection. With this awareness for the need foradequate drug therapy, a corresponding awareness has developed that suchadequate drug therapy is difficult to provide.

For example, patients with urinary tract infections often have anassociated condition of stasis, stone or obstruction in the urinarytract and the chronicity or recurrence of the urinary tract infectionsrenders unlikely successful treatment. Where such conditions exist andwhere the infection reoccurs, antibiotics often lose their effect due tothe rapid development or acquisition of resistant mutant organisms.Still further, when a patient has a urinary stone, while the infectionmight be satisfactorily treated initially with a conventionalantiobiotic, the stone contains viable bacteria which serve as a sourcefor reinfection. Accordingly, while conventional antibiotics may provideshort time relief from urinary tract infections complete cure freuentlycannot be achieved through the administration of the conventionalantibiotic agent. However, with synthetic anti-bacterial agents, thedevelopment of resistance is much less common.

Based upon the effectiveness of synthetic anti-bacterial agents in thetreatment of urinary tract infections, methenamine has had a traditionalrole for some time in this treatment. Methenamine was first synthesizedin 1860 and the use of methenamine in the treatment of cystitis wasreported as early at 1894. The methenamine exerts its anti-bacterialeffect in an acid medium by releasing formaldehyde in a concentrationwhich is bacteriostatic or bactericidal to virtually all known bacteria.

Methenamine, hexamethenamine tetramine, a tertiary amine, has propertiesof a monoacidic base in its salt formation. As a result different formsof methenamine have been developed and made available as a salt ofmethenamine and a pharmacologically aceptable organic acid. These formswere developed since they presumably acidify the urine, therebyenhancing the effect of methenamine, which can effectively release theactive formaldehyde only in an acid urine. In addition, someanti-bacterial action for the organic acids themselves has beendescribed; however, these agents have proven to be ineffective inacidifying urine in the presence of infection of the urinary tractassociated with urea-spliting pathogens. In this respect nearly allspecies of Proteus and a number of strains of Pseudomonas, Klebsiella,E. Coli and Staphlococcus produce urease, an enzyme which splits theurea according to the following reaction:

    NH.sub.2 CONH.sub.2 +  2H.sub.2 O urease 2NH.sub.3 + H.sub.2 CO.sub.3

    2NH.sub.3 + H.sub.2 O → NH.sub.4 + OH.sup.-

the hyperammonuria and alkalinity which result through the foregoingreaction appear to be necessary for the precipitation of struvite (MgNH₄PO₄.6 H₂ O), the predominant component of infected urinary calculi. Thisalkalinity eliminates the anti-bacterial activity of methenamine andcompounds based on methenamine, thereby making these materialsineffective anti-bacterial agents for treatment of infection due tourease producing uropathogens when used in typical anti-bacterialdosages.

Due to the foregoing, where a patient is suffering from a urinary tractinfection associated with certain urease producing bacteria, especiallyspecies of Proteus, the commonly employed anti-bacterial agentmethanamine and compounds based on methenamine are ineffective in thetreatment of the infection due to the hyperammonuria and alkalinitywhich result from the urease producing bacteria. Such alkalinityprevents the conversion of methanamine to its active anti-bacterial fromformaldehyde. Based upon the foregoing, the art has long sought a methodof treating urinary tract infections and a composition for use in suchmethod which can eliminate the foregoing problem by maintainingphysiologic acidity of urine and thereby provide the necessaryenvironment for effective methenamine conversion to formaldehyde, evenin the presence of urease producing bacteria.

SUMMARY OF THE INVENTION

It has now been discovered in accordance with the present invention thatthe foregoing disadvantages and drawbacks of previously proposedprocedures for treatment of urinary tract infections can be eliminatedthrough the provision of the method and composition of the presentinvention whereby a patient suffering from urinary tract infection,especially an infection caused by bacteria of the species Proteus isadministered an effective anti-bacterial dosage of a composition whichcomprises in combination (a) a source of methanamine, e.g., methenamineand (b) a source of hydroxamate groups, e.g., acetohydroxamic acid, thesource of hydroxamate groups being administered in an amount effectiveto potentiate or synergize the anti-bacterial effect of the source ofmethenamine by preventing the alkalinization normally induced by ureaseproducing bacteria.

In connection with the above it has been discovered in accordance withthe present invention that the administration of a source of hydroxamategroups, perferably acetohydroxamic acid in combination with a source ofmethenamine allows the methanamine to act as an effective anti-bacterialagent in the treatment of a urinary tract infection in that the sourceof hydroxamate groups as a urease inhibitor eliminates thealkalinization associated with the bacterial urease invariably producedby Proteus and some other species of bacteria, which alkalinization hasheretofore eliminated the effectiveness of the methenamine.

Potentiation of the anti-bacterial effect of a source of methenamine bya source of hydroxamate groups is observed in accordance with thepresent invention at a urinary concentration of methenamine of fromabout 0.05 to about 1.0 mg/ml and a urinary concentration of hydroxamategroups of from about 0.05 to about 2.0 mg/ml.

In addition, it has now been discovered in accordance with the presentinvention that the urinary pathogenicity of urease producing bacteriainduced infection can be eliminated by administering to a patientsuffering from such infection an effective amount of a source ofhydroxamate groups, e.g., acetohydroxamic acid. This result which isachieved in accordance with the present invention is predicated upon thediscovery that the pathogenicity of the infection is associated with theurease production, the inhibiting of which, by the administration of aurease inhibitor, can effectively eliminate the pathogenicity of thebacteria, while not affecting the colonization of the bacteria. Againhowever, the pathogenicity and colonization can be eliminated by theconjoint administration of both the source of methenamine and the sourceof hydroxamate groups.

In has been further discovered in accordance with the present inventionthat urinary stones, e.g., struvite and apatite stones generallyassociated with urease producing bacteria can be effectively dissolvedby administering a source of hydroxamate groups, e.g., acetohydroxamicacid, which source of hydroxamate groups has the effect of bringingabout a physiological pH in the urine, with under-saturation withrespect to struvite and apatite crystals.

When a source of hydroxamate groups is administered alone in accordancewith the present invention a therapeutic urinary concentration ofhydroxamate groups of from about 0.05 to about 2.0 mg/ml has been foundto be very effective.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a plot of pH vs. time in hours illustrating the effectivenessof the combination of methenamine and acetohydroxamic acid on thealkalinization of urine by the growth of Proteus retteri;

FIG. 2 is a plot of viable bacteria per milliliter vs. time in hoursillustrating the synergistic anti-bacterial effect of methenamine andacetohydroxamic acid on Proteus rettgeri;

FIG. 3 is a plot of bacteria per milliliter and pH vs. time in hoursillustrating the effect of methanamine and acetohydroxamic acid in adynamic study with Proteus rettgeri;

FIGS. 4-15 are plots of time in hours vs. colony forming units permilliliter for Experiments A-L, respectively of Example 3, illustratingthe synergistic effect of acetohydroxamic acid on methenamine in astatic-dynamic study with Proteus mirabilis, Proteus rettgeri andProteus morgani;

FIG. 16 is a plot of time in hours vs. colony forming units permilliliter for a typical dynamic system experiment with Proteusmarabilis; and

FIG. 17 is a cumulative plot of methenamine concentration vs. colonyforming units per milliter for six dynamic system equipment with Proteusmirabilis.

DESCRIPTION OF PREFERRED EMBODIMENTS Elimination of Colonization andPathogenicity of Urease Producing Bacteria Induced Urinary TractInfection

The method and composition of the present invention achieve the desiredadvantages over previous methods and compositions through the provisionof a synergistic anti-bacterial effect exhibit with regard to ureaseproducing bacteria through the concurrent administration of a source ofmethenamine and a source of hydroxamate groups, the source ofhydroxamate groups being administered in an amount effective to maintainthe necessary urine acidity to allow the anti-bacterial effect of thesource of methenamine. The method of the present invention is applied tothe treatment of patients with urinary tract infections, particularlythose urinary tract infections caused by urease producing bacteria,especially those of the species Proteus.

A number of Proteus species, some of which are resistant to the highestachievable urinary concentrations of currently utilized antibiotics,have recently emerged as common hospital acquired urinary pathogens.While it has been shown that such Proteus species are susceptible totreatment with methenamine, the use of methenamine alone as ananti-bacterial agent in the treatment of urinary tract infectionassociated with such urease producing species has not been successful inthat (1) methenamine when administered in a typical anti-bacterialdosage is not effective at an alkaline pH and the urease producingspecies belong to that class of bacteria which produce an alkalinizationof the urine; (2) attempts to acidify the urine in the face ofpersisting infection associated with urease producing bacteria have notbeen successful; and (3) organic acids salts of methenamine, such asmethenamine mandelate and methenamine hippurate have no real effect onovercoming the alkalinization of the urine in the face of urinaryinfections produced by urease producing bacteria.

As indicated above, the method and composition of the present inventionare particularly applicable to the treatment of urinary tract infectionscaused by bacteria of the species Proteus. Representative examples ofsuch Proteus species include, for example, mirabilis, vulgaris, morganiand rettgeri, all of which species of Proteus are urease producingbacteria which reduce the effective anti-bacterial action of methenamineand methenamine based compounds, when such compounds are utilized fortheir anti-bacterial effect in the treatment of urinary tractinfections.

Source of Methenamine

The source of methenamine which is utilized as the primaryanti-bacterial agent in the method and composition of the presentinvention can be methenamine itself or any of those compounds whichcombine the active methenamine nucleus with additional materials,principally formed from methenamine and an acid, especially an organicacid. In this respect, any methenamine salt with a pharmacologicallyacceptable acid can be employed in accordance with the presentinvention. For example, the active methenamine can be formulated asmethenamine hippurate, methenamine mandelate or methenaminesulfosalicylate, effective urinary anti-bacterial agents. Methenaminehippurate is available from Merrell-National Labratories Division ofRichard-Merrell of Cincinati, Ohio, under the name "Hiprex" and underthe name "Urex" from Riker Laboratories, Inc. a subsidiary of 3MCompany, Northridge, Calif. Similarly, methenamine mandelate isavailable from Warner-Chilcote Laboratories, Div. Warner-LambertCompany, Morris Plains, N.J. under the name "Mandelamine" methenaminesulfosalicylate is available under the name "Hexalet". Any of theseforms as well as other forms of methenamine which contain the activemethenamine nucleus can be utilized in accordance with the presentinvention to provide the required source of methenamine.

It should be apparent from the above that the examples set forth aremerely exemplary of the forms which can be taken by the source ofmethenamine in accordance with the present invention. In this regard itis merely necessary in accordance with the present invention that theactive methenamine be administered so as to allow the methenamine toexhibit its anti-bacterial effect with regard to the urinary tractinfections. Accordingly, the expression "source of methenamine" asemployed throughout is meant to embrace methenamine as well as all ofthe forms of methenamine which can effectively provide the necessarymethenamine and its anti-bacterial effect.

Source of Hydroxamate Groups

Compounds which provide the active hydroxamate groups have been knownprior to the present invention as specific and effective inhibitors ofbacterial urease. In this regard the terminal ##STR1## group whichcharacterizes the group of compounds applicable in accordance with thepresent invention is the active site of urease inhibition and it ishypothesized that in this urease inhibition reaction two moles of thehydroxamic acid bind with one mole of urease.

Of the various compounds which contain the hydroxamate group, the foursimplest aliphatic hydroxamic acids appear to possess the greatesturease inhibiting effect. These aliphatic hydroxamic acids includingformohydroxamic acid, acetohydroxamic acid, propionohydroxamic acid andisobutyhydroxamic acid. Of these, acetohydroxamic acid is particularlypreferred. This source of hydroxamate groups has the followingcharacteristics:

                  Table 1                                                         ______________________________________                                        Molecular Wt.                75.068                                            Structure                                                                                     ##STR2##                                                     Molar Absorbancy Index                                                         in 2% FeCl.sub.3 /0.        In HCL 915                                       pKa                           9.32                                            Melting Point                89-91° C                                  ______________________________________                                    

In addition to such aliphatic hydroxamic acids, other aliphatichydroxamic acids as well as aromatic hydroxamic acids and araliphatichydroxamic acids show the required urease inhibiting effect to beapplicably employed in the method and composition of the presentinvention. Typical sources of hydroxamate groups falling within theforegoing classes are illustrated in an article "Effect of Acyl Residuesof Hydroxamic Acids on Urease Inhibition," by K. Kobashi, Kenji Kumakiand Junichi Hase, Biochem, Biophys, Acta, 227 (1971) pages 429-441. Anyof the sources of hydroxamate groups set forth in such article can besuccessfully employed in the method and composition of the presentinvention and the contents of such article are herein incorporated byreference. Accordingly, as used throughout, the expression "source ofhydroxamate groups" is meant to embrace not only the preferred materialsillustrated above, but in addition any of the conventional sources ofhydroxamate groups which prossess the necessary urease inhibitingcharacteristics.

As will be appreciated hereinafter the source of hydroxamate groupsutilized in accordance with the present invention, and acetohydroxamicacid in particular, show a slight anti-bacterial effect with regard tobacteria of the species Proteus; however, the anti-bacterial effectwhich is shown through the combination of a source of methenamine and asource of hydroxamate groups is substantially in excess of thearithmetic sum of the anti-bacterial effects of the individualcompounds. Accordingly through the provision of the combination of thesource of methenamine and source of hydroxamate groups a synergisticanti-bacterial effect has been created, which synergistic anti-bacterialeffect allows the treatment of urinary tract infections, specificallythose caused by bacteria of the species Proteus in a manner notheretofore possible with conventional treatments and compositions.

While the precise mechanism for the synergistic potentiation of theanti-bacterial effect of the sources of methemanine is not known, itappears as though this potentiation is associated with the ureaseinhibiting characteristic of the source of hydroxamate groups, whichurease inhibiting characteristic eliminates the alkalinization of theurine associated with infections caused by bacteria of the speciesProteus. In this regard since methenamine and compounds based onmethenamine depend upon acidic pH and the effective anti-bacterialactivity thereof is eliminated in an alkaline medium, the elimination ofthe alkalinization through the conjoint utilization of the source ofhydroxamate groups allows the pH of the urine to be maintained in thatrange in which the source of methenamine is most effective as ananti-bacterial agent.

While some potentiation of the anti-bacterial effect of the source ofmethenamine can be explained through the urease inhibiting effect of thesource of hydroxamate groups and the inhibition of the alkalinization ofthe urine associated with the bacteria of the species Proteus, it shouldbe recognized that a truly unexpected synergistic effect is achieved.

Dosages and Urine Concentrations

The anti-bacterial effect of methenamine and methenamine based compoundsis apparently associated with the release of formaldehyde, with theformaldehyde formed being the active anti-bacterial agent. Accordingly,the effective dosage and concentration of the source of methenamine inaccordance with the present invention can be defined as that dosage orconcentration which provides the desirable anti-bacterial level offormaldehyde in the urine. No precise optimum concentration offormaldehyde in the urine resulting from the release of formaldehydefrom methenamine or methenamine based compound can be set forth sincethis will vary depending upon many factors, including the degree ofinfection and the particular bacterium of the species causing theinfection. However, it has been discovered in accordance with thepresent invention that an effective anti-bacterial level of formaldehydein the urine is achieved with a urinary concentration of methenaminewithin the range of about 0.05 to about 1.0 milligram per milliliter.

The particular dosage of methenamine or methenamine based compound toachieve the foregoing concentration of methenamine in the urine willdepend upon many factors, including the body weight and physiology ofthe patient, renal activity of the patient and effectiveness of thesource of methenamine to release the formaldehyde as the necessaryanti-bacterial agent, this latter factor being controlled in accordancewith the present invention by the conjoint administration of a source ofhydroxamate groups. While the foregoing is true, generally the desiredconcentration of methenamine in the urine can be achieved byadministering an oral dosage of methenamine or methenamine basedcompound of about 0.5 to about 8 grams per day, preferably 1 to 4 gramsper day. It should be recognized that while such guidelines are setforth, the present invention is not in any way limited thereto sinceslightly greater dosages may be required in particular cases andslightly lesser dosages may be required, such as in the case of anadministration of the source of methenamine and source of hydroxmategroups to children and infants. Generally, however, the desiredconcentration of methenamine in the urine can be achieved byadministration of the above dosage.

As previously indicated, all that is necessary in accordance with thepresent invention to achieve the desired result is that the source ofhydroxamate groups be administered in an amount effective to provide asynergistic effect with regard to activity of the source of methenamine.Here again, the dosage can be defined in terms of the desired urinaryconcentration of the hydroxamate groups, with the desired concentrationbeing from about 0.05 to about 2.0 milligrams per milliliter. It is ofcourse noted that within the foregong guidelines, dosages near the upperend of the range may be required in the initial treatment of theinfection whereas dosages near the lower end of the range will besatisfactory to maintain the desired treatment. When the desired urinaryconcentrations are converted to oral dosages of the source ofhydroxamate groups, it is generally found that a dosage of about 0.25 toabout 4 grams per day of the source of hydroxamate groups will achievethe desired urinary concentration. Again, however, these dosages aremerely set forth as guidelines and slightly greater and slightly lesserdosages may be administered where desired for particular purposes orwhere required based upon the physiology of the patient.

With regard to the ratio of the source of methenamine to the source ofhydroxamate groups, no critically exists in accordance with the presentinvention as long as the individual components are employed in theamounts set forth above, i.e., the source of methenamine is employed inan amount effective to show the desired anti-bacterial effect and thesource of hydroxamate groups is employed in an amount effective toprovide a synergistic effect with respect to the anti-bacterial activityof the source of methenamine and to exhibit the desired ureaseinhibition. Accordingly, within these guidelines operable dosages andamounts can be selected for any particular case.

The sensitivity of methenamine and methenamine based compounds to pH iswell known. In this regard methenamine for example, shows excellentrelease of formaldehyde at a pH of 5.0 and good release of formaldehydeat a pH of 5.5. At a pH of 6.0 the release of formaldehyde is generallyadequate for the treatment of most urinary tract infections, but as thepH of the urine reaches 6.5 insufficient conversion and release offormaldehyde is achieved and the anti-bacterial effect of themethenamine is totally diminished. Because of this problem, the art haslong sought a mechanism by which the effective anti-bacterialcharacteristics of methenamine can be achieved even in the face ofurease producing bacteria, this now being achieved through the provisionof the present invention comprising the conjoint administration of asource of methenamine and a source of hydroxamate groups.

The form of the compounds administered in accordance with the presentinvention can be any of the conventially utilized for theadministratiion of pharmacologically effective compounds, orally orparenterally. Thus, for example, the source of methenamine and thesource of hydroxamate groups can be administered in a liquid formutilizing a conventional alcohol or similar vehicle or alternatively thesource of methenamine and source of hydroxamate groups can beadministered in the form of tablets or in the form of capsules,utilizing conventional fillers, excipients, etc. to provide thenecessary form of the desired dosages.

Elimination of Pathogenicity of Urease Producing Bacteria InducedUrinary Tract Infections

Various investigators have put forward the hypothesis that urease isresponsible for the ability of Proteus and similar bacteria which areurease producing bacteria to cause urinary tract infections. Thishypothesis is supported by the fact that bacteria of the Proteus speciesrarely cause infections outside of the urinary tract, although they arevery frequent pathogens in the urinary tract. This is apparently due tothe fact that urine is the only substance in the body that containssubstantial amounts of urea, i.e., sufficient urea that when broken downby urease a sufficient amount of ammonia is generated to causealkalinization. The pathogenicity as referred to herein relates to thetissue damage that results when urease producing bacteria infect theurinary tract and is distinguishable from the colonization by thebacteria which can exist without damage to tissues.

Since infection can be defined as damage to tissue by micro-organismsand always implies some host response, such as an outpouring of whiteblood cells, unless of course the host has been pretreated in a mannerto prevent this response, the elimination of the pathogenicity of theinfection can effectively eliminate the infection itself while allowingthe colonization of the bacteria, which colonization does not itselfcause tissue damage. This has been achieved in accordance with thepresent invention by administering the urease inhibiting compoundspreviously described, i.e., source of hydroxamate groups, andacetohydroxamic acid in particular. These compounds effectively inhibitthe urease which confers pathogenicity of the Proteus species ofbacteria. By inhibiting the urease in this manner and by eliminating thepathogenicity of the infection, the infection itself is eliminated andtissue damage prevented, though colonization of the bacteria maypersist. This is achieved in accordance with the present inventionwithout the simultaneous addition of an anti-bacterial agent, such as asource of methenamine, which is effective to eliminate the colonizationof the urease producing bacteria. Accordingly, this aspect of thepresent invention provides for the administration of only the source ofhydroxamate groups, with an associated elimination of the pathogenicityof the infection, absent elimination of bacteria colonization.

As will be appreciated from the later present examples, by administeringonly the source of hydroxamate groups it is possible to effectivelyeliminate tissue damage caused by urease producing bacteria byinhibiting the action of the pathogenic factor, namely urease,notwithstanding the fact that colonization of the bacteria is noteliminated. In this respect the colonization of the bacteria whichcontinues does not produce adverse effects, such as tissue damage, sincethe pathogenic agent responsible for the infection and associated tissuedamage is eliminated. Accordingly, the present invention, byadministration of only the active urease inhibiting compound, i.e.,source of hydroxamate groups, is a simple and efficient means oftreating and eliminating urinary tract infections, specifically thoseurinary tract infections associated with urease producing bacteria,especially those of the species Proteus.

The foregoing discussion involves a unique approach to the treatment ofinfections in that the approach does not necessarily aim towardelimination of the bacteria with which the infection is associated butrather toward blocking the effect of the pathogenic agent produced bythe bacteria, whereby the tissue damage induced by the bacteria and/orbacterial agent is stopped. This phenomenon provides a new simplifiedapproach to the treatment of infection.

In connection with this approach to the treatment of infections it ispointed out that the dosages and forms of the active urease inhibitingcompound, i.e., source of hydroxamate groups are the same as previouslyset forth in connection with the combined administration of a source ofhydroxamate groups and a source of methenamine. The amount ofhydroxamate groups introduced into the system should be that amounteffective to produce the desired urease inhibiting effect, such that theactive pathogenic agent, i.e., urease can be inhibited and tissuedamages associated with this pathogenic agent stopped.

Stone Dissolution

It is a general contention that bacterial infection of the urine maycause urinary stone formation and there is considerable dataestablishing the cause and effect relationship between urea-splittingurinary infections and struvite and calcium apatite stone formation. Itis hypothesized that urease producing bacteria cause stone formation dueto the alkalinity produced by the hydrolysis of urea. Consideration hasbeen given to dissolving of the urinary stones since, as previouslynoted, the bacteria within the stone can reinfect the urinary tract.

It has been discovered in accordance with the present invention thaturease producing bacteria promote stone formation by way of the enzymeurease. Moreover, it has been discovered in accordance with the presentinvention that by administering to a person suffering from infectedurinary stones an effective urease inhibiting amount of an inhibitor,such as the sources of hydroxamate groups previously described, andacetohydroxamic acid in particular, it is possible to maintain the urineat a physiological pH and thereby produce an environment effective forthe dissolution of urinary stones.

Sterile urine from patients not suffering from urinary tract infectionsis usually acidic and under-saturated with respect to struvite andapatite crystals. Due to such undersaturation such urine will dissolvestruvite and apatite stones. By preventing alkalinization andmaintaining urine in a state of under-saturation, despite the presenceof urease producing bacteria, it is possible in accordance with thepresent invention to provide a urine with a physiological pH that willdissolve such struvite and apatite crystals in the form of urinary tractstones. Accordingly, the method of the present invention involvesadministering an effective urease inhibiting amount of a source ofhydroxamate groups to a patent suffering infection from urinary tractstones for the purpose of dissolving such stones.

With respect to this embodiment of the present invention the types ofcompound effective to dissolve the stones are the same compoundspreviously described. In addition, the dosages set forth previously withregard to such introduction of a source of hydroxamate groups areessentially the same as employed in this embodiment of the presentinvention with respect to dissolving struvite and apatite stones. Inthis embodiment the source of hydroxamate groups can be administeredeither alone or in combination with a source of methenamine to bothdissolve the stones and eliminate the colonization of the ureaseproducing bacteria.

The methods and composition of the present invention will now beillustrated by way of specific examples. It should be recognized thatthese examples are presented solely for the purpose of illustration andthe present invention cannot under any circumstances be deemed limitedthereby.

EXAMPLE 1

Static studies were conducted to illustrate the synergistic effectachieved in accordance with the present invention through the combinedemployment of a source of methenamine, i.e., methenamine itself and asource of hydroxamate groups, i.e., acetohydroxamic acid.

In order to achieve reproducibility of results with large quantities orurine required for these studies a synthetic urine was formulated basedon the proportion of solutes found in the urine from normal humansubjects. The composition of such urine was as follows:

    ______________________________________                                        COMPOSITION OF SYNTHETIC URINE                                                ______________________________________                                        Compound      Gm/L        Concentration/L                                     ______________________________________                                        CaCl.sub.2 . 2H.sub.2 O                                                                     0.65        Ca - 4.3mM                                          MgCl.sub.2 . 6H.sub.2 O                                                                     0.651       Mg - 3.2mM                                          NaCl          4.6                                                             Na.sub.2 SO.sub.4                                                                           2.3         SO.sub.4 - 16mM                                     Na.sub.3 Citrate . 2H.sub.2 O                                                               0.65        Citrate - 2.3mM                                     Na.sub.2 Oxalate                                                                            0.020       Oxalate - .149mM                                    KH.sub.2 PO.sub.4                                                                           2.8         PO.sub.2 - 20.5mM                                   KCl           1.6                                                             NH.sub.4 Cl   1.0         NH.sub.4 - 19mEq                                    Urea          25.0                                                            Creatinine    1.1                                                                                       Total Na = 118mEq                                                             Total K =  42mEq                                    ______________________________________                                    

Brain heart infusion (BHI) broth in a final concentration of 5% wasadded to support bacterial growth. The pH of the synthetic urine priorto addition of the BHI broth was 5.7 - 5.8. It was found that the growthof the Proteus bacteria in the synthetic urine an the alkalinization ofthe synthetic urine utilized proceeded at approximately the same rate asin random human urine from normal subjects.

In these tests the methenamine was utilized in a final concentration of0.9 milligrams per milliliter. Acetohydroxamic acid was synthesized bythe procedure set forth in "PREPARATION AND SOME PROPERTIES OF STABLEAND CARBON -14 AND TRITIUM-LABELED SHORT-CHAIN ALIPHATIC HYROXAMICACIDS", Fishbein, W. N., J. J. Dally and C. L. Streeter, 1969,Analytical BioChem 28:13-21. The acetohydroxamic acid which wassynthesized was added to the urine to provide a concentration of 1.9mg/ml. These concentrations which were selected in the static studieswere concentrations which would be achievable in the urine of patientswith normal renal function.

In accordance with these studies methenamine alone, the acetohydroxamicacid alone and both methenamine and acetohydroxamic acid were added tothe synthetic urine in the above concentrations and aliquots of thethree types of samples, as well as the synergistic urine alone, wereadjusted at 0.1 pH unit intervals from 5.6 to 6.1 by titration with 1 NHCl or 1 N NaOH. At each pH level a set of four tubes was obtained, withthese tubes being as follows:

1. Urine alone (control);

2. Urine plus methenamine;

3. Urine plus acetohydroxamic acid; and

4. Urine plus acetohyroxamic acid plus methenamine.

Bacteria of the species Proteus were added to yeild 10⁷ organisms permilliliter and each of the tubes was incubated at 37° centigrade. Atvarying time intervals aliquots were removed for pH determination andbacterial enumeration by serial tenfold dilution.

As a result of such static tests is was found that the growth of theProteus bacteria in the synthetic urine alone caused the pH to rise togreater that pH 8.0 within 8 hours. This illustrates the effect ofurease production by Proteus bacteria on the pH of urine. It was furtherfound that the rate of alkalinization varied with the bacterial strainbut was unrelated to the initial pH.

With respect to the samples containing synthetic urine and theacetohydroxamic acid it was found that the acetohydroxamic acidinhibited alkalinizaton for the first eight hours, but its effects as aurease inhibitor were overcome within 24 hours by bacterial poliferationleading to an alkaline pH. With regard to the samples containingsynthetic urine and methenamine it was found that the anti-bacterialeffect of methenamine was dependent upon the starting pH. When theinitial pH was less than 5.3 only a slight upward drift was observed,consistent with secondary metabolic reactions in dead and dyingbacterial cells. At a pH greater than 6.5 the presence of methenaminedid not alter progressive alkalinization. Between these extremesalkalinization was inhibited in proportion to the initial urine acidity.When methenamine and acetohydroxamic acid were combined in the syntheticurine, a synergistic effect was exhibited, with alkalinization beinginhibited to approximately the same degree between pH 5.5 and 6.1 withall of the Proteus species studied, i.e., Proteus mirabilis, Proteusvulgaris, Proteus morgani and Proteus rettgeri. The synergistic effecton the alkalinization of urine by the growth of Proteus rettgeri can beseen in FIG. 1.

With regard to the alkalinization of urine by the growth of Proteusrettgeri a synergistic effect is achieved when methenamine andacetohydroxamic acid are combined in the synthetic urine, in thatalkalinization is inhibited through the combination to about the samedegree between pH 5.1 and 6.1 and no undesirable alkalinization is seen.This is compared to the case of urine plus methenamine alone and urineplus acetohydroxamic acid alone.

The synergistic effect achieved in accordance with the present inventionwith regard to bacterial growth can be seen by reference to FIG. 2 whichplots the viable bacteria per milliliter against time in hours, thebacteria being Proteus rettgeri. Results similar to those of FIG. 2 wereseen for the other Proteus species studied.

As seen from FIG. 2 bacterial counts in urine rose to greater than 10⁸per milliliter within four hours. The acetohydroxamic acid exerted aslight bacteriostatic effect; however, within eight to eighteen hoursthe number of bacteria approached 10⁸ per milliliter. The anti-bacterialeffect on the methenamine was proportional to the initial pH and asillustrated in FIG. 2 the acetohydroxamic acid potentiated thebactericidal effect of methenamine at every pH between 5.5 and 6.1 andthis synergistic effect was seen for all Proteus species studied. It canbe seen from FIG. 2 that the bacterial effect achieved through thecombined presence of methenamine and acetohydroxamic acid is far greaterthan the bacterial effects shown for methenamine alone, acetohydroxamicacid alone or what would be expected from the arithmetic sum of the two.

EXAMPLE 2

Dynamic studies were conducted to further illustrate the synergisticeffect associated with the combined presence of a source of methenamine,i.e., methenamine itself and a source of hyroxamate groups, i.e.,acetohydroxamic acid in the urine.

Utilizing the same synergistic urine as in example 1, a dynamic systemwas designated to resemble a Proteus infected human bladder. Utilizing amulticassetts roller pump, urine of a pH of 5.85 was pumped at 40milliliters per hour from three reservoir flasks into three reactionchambers. The synthetic urine in flask 1 contained no acetohydroxamicacid, while the urine in flask 2 and 3 contained 1.9 milligramsacetohydroxamic acid per milliliter. An aqueous solution of 18milligrams per milliliter of methenamine at a pH of 7.5 was pumped intoreaction chambers 1 and 3 at a rate of two milliliters per hour yieldinga final concentration of methenamine of 0.9 milligrams per milliliter.As a result of this reaction chamber 1 contained methenamine, reactionchamber 2 contained acetohydroxamic acid and reaction chamber 3contained methenamine plus acetohydroxamic acid. A control containingurine alone was not carried out in the dynamic study since preliminaryinvestigations had shown that the 0.9 milligrams per millimetermethenamine affected growth of Proteus and alkalinization of urine onlyslightly in this dynamic system when compared to the synthetic urinealone. The reaction chambers contained gently rotating magnetic stirrersand were kept in a warm bath at 37° C.

At the start of this dynamic experiment 5 milliliters of urine at a pHof 8.9 containing 10⁷ Proteus per milliliter were added to each reactionchamber. At 4 - 6 hour intervals the volume of urine in each reactionchamber was recorded, the pH of each reaction chamber was determined andserial dilutions were carried out for bacterial enumeration. Thereaction chambers were then emptied leaving a residual urine volume of 5milliliters.

The results which were achieved in this dynamic test are illustrated inFIG. 3, which sets forth the results with regard to the dynamic studywith Proteus rettgeri. Similar results were achieved with each of theProteus bacteria studied.

As seen in FIG. 3, in the urine containing methenamine, at the end ofeach four to eight hour period the pH always exceeded 8.5, i.e., a pH atwhich there is no anti-bacterial effect of methenamine. The number ofbacteria per milliliter fell slightly during the first six hours, but bythe end of the second time period the colony count had risen to greaterthan 10⁷ per milliliter where it remained throughout the study. However,in the presence of acetohydroxamic acid alkalinization of Proteus wasinhibited and the pH fell to 5.85 - 6.1 within the first six hours andremained in that range throughout the study. The acetohydroxamic acidexhibits a minor bacteriostatic effect maintaining the number ofbacteria between 10⁵ and 10⁷ per milliliter. In the presence of both theacetohydroxamic acid and methenamine, the pH remained acid throughoutand there was a steady decline in colony count resulting insterilization of the system within 24 - 48 hours. This illustrates thatthe acetohydroxamic acid and methenamine together exert a synergisticeffect against urease producing bacteria, especially those of theProteus species.

EXAMPLE 3

In this example in vitro investigations were carried out using astatic-dynamic system delineating (1) bacteriostatic dose-responserelation for methenamine against Proteus, and (2) a synergisticbacteriostatic dose-response relation for methenamine andacetohydroxamic acid against Proteus.

Experimental Model

This experiment model to be described seeks to demonstrate sequentialdilution of growth inhibited bacteria at 6 hour intervals.

Tubes containing 10 ml synthetic urine was inoculated with Proteusmirabilis, Proteus rettgeri or Proteus morgani to give a final bacterialconcentration of 10⁴ -10⁷ per ml. Separate tubes contained (1) urineonly, (2) urine and acetohydroxamic acid, and (3) urine and methenamine,(4) urine, acetohydroxamic acid and methenamine. The tubes were placedin a bacteriological incubator at 37° C. At 6 hour intervals 100microliters of urine were withdrawn from each tube and the tube wasdiscarded. The 100 microliter aliquot was added to 10 ml of syntheticurine which contained the same concentration of constituents (exceptbacteria) as the initial tubes. Incubation of the tubes at 37° andsequential dilutions were made a 6 hour intervals for 24 hours. Thevariables for each of eight experiments are detailed in Table 2.

                                      TABLE 2                                     __________________________________________________________________________    EXPERIMENTAL VARIABLES                                                        __________________________________________________________________________                Initial Bacterial                                                             Inoculum          AHA conc.*                                                                           No. 1 Meth. conc.*                                                                      No. 2 Meth. conc.*             Expt.                                                                             Bacteria                                                                              cfu/ml     Urine pH*                                                                            mg/ml  mg/ml     mg/ml                          __________________________________________________________________________    A   P. mirabilis                                                                          5 × 10.sup.4                                                                       5.9    0.2    0.05      0.1                            B   P. mirabilis                                                                          3 × 10.sup.5                                                                       5.9    0.2    0.05      0.1                            C   P. mirabilis                                                                          3 × 10.sup.6                                                                       5.9    0.2    0.05      0.1                            D   P. mirabilis                                                                          5 × 10.sup.7                                                                       5.9    0.2    0.05      0.1                            E   P. mirabilis                                                                          5 × 10.sup.6                                                                       5.9    0.2    0.1       0.2                            F   P. mirabilis                                                                          8 × 10.sup.7                                                                       5.9    0.2    0.1       0.2                            G   P. mirabilis                                                                          3 × 10.sup.6                                                                       5.9    0.1    0.05      0.1                            H   P. mirabilis                                                                          6 × 10.sup.7                                                                       5.9    0.1    0.05      0.1                            I   P. mirabilis                                                                          5 × 10.sup.7                                                                       6.5    0.1, 0.05                                                                            1.0       0.5                            J   P. mirabilis                                                                          1 × 10.sup.8                                                                       7.0    0.1, 0.05                                                                            1.0       0.5, 0.2                       K   P. rettgeri                                                                           7 ×  10.sup.7                                                                      5.9     0.05  0.05      0.1                            L   P. morgani                                                                            1 × 10.sup.8                                                                       5.9    0.1, 0.05                                                                            0.05, 0.01                                                                              0.1                            __________________________________________________________________________     *The initial pH and concentration of constitutents was identical in all       tubes at the beginning of each six hour incubation.                      

Bacterial colony forming units (cfu) and urine pH were determined at thebeginning and at the completion of each six hour incubation. The cfuunits present after six hours of incubation are graphed in FIGS. 4-15.Bacteriostasis was demonstrated by a decline in the number of cfu aftereach incubation. When bacteriostatis was absent, the number of cfu aftersix hours of incubation was equal to or greater than the number of cfuat the beginning of incubation.

Bacteria

A strain of Proteus mirabilis isolated from the urine of patients withurinary tract infections was used in most experiments. Overnight growthin brain heart infusion (BHI) broth regularly produced 10⁹ cfu/ml. Theovernight culture was centrifuged at 3000 rpm for 20 minutes. Thesupernatant was discharged, and the bacterial pellet was resuspended inan equal volume of synthetic urine. The bacteria were resuspended insynthetic urine to prevent differences in pH as the bacteria wereinoculated into tubes of synthetic urine. The bacterial suspension wasinoculated into 10 ml of synthetic urine to achieve final concentrationsof 10⁴ - 10⁷ cfu/ml. Bacteria were counted by making serial ten-folddilutions following which 0.1 ml aliquots were streaked onto MacConkey'sagar.

Urine

The urine utilized was a synthetic urine as illustrated in Example 1.

Methenamine and Acetohydroxamic Acid

Methenamine was used in final concentrations from 0.05 mg/ml to 0.20mg/ml. Acetohydroxamic acid was synthesized in the same manner asdescribed in Example 1 and was added to give final concentrations of0.05-0.20 mg/ml. The results are illustrated in FIGS. 4-15 which relatesrespectively to Experiment A-L of Table 2. Each of FIGS. 4-15 representscolony forming units (cfu) present at the completion of six hourincubations. 100-fold dilutions were made prior to initiating subsequentsix hour incubations. Each figure also includes a line representative oftheoretically perfect bacteriostasis.

When bacteriostatis was absent, bacteria grew rapidly, usually reachingequal to or greater than 10⁸ per ml within the first six hours ofincubation. Significant bacteriostasis resulted in a reduced bacterialcount (cfu) with each subsequent dilution.

Acetohydroxamic Acid Only

Acetohydroxamic acid at a concentration of ≦0.9 mg/ml exerted nobacteriostatic effect. The acetohydroxamic acid data was omitted fromTable 2 and FIGS. 4-15 in an effort to simplify the presentation.

Methenamine Only

Experiments A and B suggest that 0.05 mg/ml methenamine has somebacteriostatic activity in urine at pH 5.9 against small concentrationsof Proteus mirabilis (10⁴ /ml and 10⁵ /ml) (FIGS. 4, 5). Experiments Cthrough H demonstrate no bacteriostasis by 0.05 mg/ml methenamineagainst larger concentrations of bacteria (≧10⁶ /ml Proteus).

All experiments performed in acidic urine demonstrate bacteriostasis by0.1 mg/ml methenamine (FIGS. 4-7, 10 and 11. Experiments E and F (FIGS.8, 9) demonstrate bacteriostasis by 0.2 mg/ml methemanine. Theseconcentrations of methenamine were bacteriostatic against allconcentrations of Proteus (10⁴ - 10⁷ per ml). Methenamine at aconcentration of 0.2 mg/ml consistently exerted a greater bacteriostaticeffect than did methenamine 0.1 mg/ml (FIGS. 8, 9).

Methenamine and Acetohydroxamic Acid

In every experiment involving acidic urine the combination ofmethenamine and acetohydroxamic acid exerted a more pronouncedbacteriostatic effect than did methenamine alone (FIGS. 4-11, 14 and15). In experiment A, 0.2 mg/ml acetohydroxamic acid enhanced the mildbacteriostatic effect of 0.05 mg/ml methenamine against smallconcentrations of bacteria. The 0.05 mg/ml methenamine had little or nobacteriostatic activity against large concentrations of bacteria (≧ 10⁶Proteus/ml), the combination of methenamine 0.05 mg/ml andacetohydroxamic acid 0.2 mg/ml did exert a reproducible bacteriostaticeffect (FIGS. 4-7).

In experiments K and L (FIGS. 14 and 15) 0.05 mg/ml acetohydroxamic acidpotentiated the bacteriostatic effect of 0.05 mg/ml methenamine.Methenamine and the acetohydroxamic acid - methenamine combination weremuch less bacteriostatic in alkaline urine (FIGS. 12 and 13) than inacidic urine.

EXAMPLE 4

This example was carried out using a dynamic model as described inconnection with Example 2. This model more closely approximates thedynamics of the urinary tract, i.e., residual urine after simulatedvoiding is approximately 5 ml and bacteria may adhere to particulatematter or to the walls of the container.

Bacteria

Proteus species, isolated from the urine of patients with known urinaryinfection, were maintained in the laboratory by monthly transfer on agarslants. These organisms grew to about 10⁹ colony forming units (cfu) perml after overnight growth at 37° C in brain heart infusion broth (BHI).Bacteria were counted by carrying out serial tenfold dilutions andplating 0.01 ml aliquots onto BHI agar (enriched with 2% additional agarto prevent bacterial swarming) and MacConkey' agar.

Synthetic urine.

Synthetic urine formulated as previously described (Ex. 1) was used. ThepH of this synthetic urine was 5.85.

Acetohydroxamic Acid

Acetohydroxamic acid was synthesized as described in Example 1; thechemical preparations used for these studies were 98-99% pure.

Ammonia determinations

The concentration of ammonia in urine was measured using an Orionammonia electrode (Orion Instruments, electrode No. 95-10).

In this dynamic system urine with or without acetohydroxamic acid waspumped at 48 ml/hr from reservoirs via an auto-analyzer pump intoreaction chambers which were incubated in a shaking water bath at 37° C.Methenamine from a separate reservoir at an alkaline pH was added at 1.8ml/hr into some of the reaction chambers to yield final concentrationsranging from 0.1 to 0.5 mg/ml. Every six hours the reaction chamberswere emptied except for a 5 ml residual, and the sequence was repeated.In these experiments the pH, ammonia concentration and cfu per ml urinewere determined for each reaction chamber at the end of each six hourperiod.

In this dynamic system acetohydroxamic acid consistently potentiated theanti-bacterial activity of methenamine. Results of a representativestudy using 0.5 mg/ml methenamine and 0.5 or 1.0 mg/ml acetohydroxamicacid are shown in FIG. 16. Methenamine alone and the higherconcentration of acetohydroxamic acic slightly inhibited bacterialgrowth. Either concentration of acetohyroxamic acid together withmethenamine caused a marked decline in cfu; as has been shown formethenamine in other systems, an actual bactericidal effect seems tohave been observed. In this study 1 mg/ml acetohydroxamic acidmaintained the pH at normal levels without methenamine being present.The initial tendency for the pH to neutralize in the presence ofmethenamine alone was overcome by the end of 18-24 hours. With lowerconcentrations of methenamine the pH rapidly became alkaline andremained so throughout. The presence of acetohydroxamic acid andmethenamine together kept the pH in a normal range. Ammoniaconcentrations closely followed pH in all the studies that were done. Inthis dynamic model synergy was observed for all concentrations ≧ 0.2mg/ml acetohydroxamic acid and ≧0.2 mg/ml methenamine. As little as 0.1mg/ml methenamine was bacteriostatic in the presence of 1.0 mg/mlacetohydroxamic acid. Cumulative results of all studies which indicatebacterial counts at the end of 18 hours in each individual experimentare shown in FIG. 17. In all of these experiments prevention ofalkalinization by acetohydroxamic acid and methenamine together could bedemonstrated with slightly lower concentrations of both compounds thanthose required to demonstrate bacteriostasis.

EXAMPLE 5

This example was carried out to illustrate the effectiveness ofadministering a source of hydroxamate groups, e.g., acetohydroxamic acidin eliminating the pathogenicity of urinary tract infections and indissolving or reducing infected bladder stones.

The formation of infected bladder stones was induced in 250 gram maleSprague-Dawley rats, according to the method described in "ExperimentalUrolithiasis VIII -- Furadantin in Treatment of Experimental ProteusInfection with Stone Formation" C. W. Vermuelen and R. Goetz, Journal ofUrology, 72:99, 1954. Rats wer anesthetized with ether and the abdominalwall swabbed with 70% ethyl alcohol. The urinary bladder was enteredthrough a small suprapubic incision. Zinc discs, 6 mm. in diameter, weredipped into a saline suspension containing 10⁷ Proteus mirabilis per ml(infective discs) and immediately inserted into the bladder. Discshandled in this fashion carried 10⁵ ± 15% organisms. The bladder wasclosed with a single purse-string suture of 3-0 chromic catgut, and theskin was approximated with metallic clips. All animals were fastedovernight following the surgical procedure and resumed their usual dietthe following day.

Rats were housed individually in stainless steel metabolic cages andwere fed Purina Rat Chow pellets ad lib. Fluid was given in stainlesssteel cups as a solution of 5-20% sucrose in water.

Treatment with acetohydroxamic acid was begun in half the rats the dayafter surgery. These animals were offered 150 mg. of acetohydroxamicacid daily in 20% sucrose-cola solution of which they ingested anaverage of 100 mg. The concentration of acetohydroxamic acid in theirurine varied from 0.3 to 7.8 mg/ml (means 2.88 mg/ml). Control animalsfor this study received no acetohydroxamic acid. Their fluid intake wasadjusted to equal that of the acetohydroxamic acid treated group by pairwatering. All animals were sacrificed on the seventh day.

The urine pH in the control animals was persistently alkaline after theProteus mirabilis urinary infection was induced. Urine cultures obtainedat autopsy grew Proteus mirabilis > 10⁵ organisms/ml and bladder stoneformation was abundant (mean 52.1 ± 27 mg) in all control rats exceptfor one, which had sterile, acidic urine. This animal appeared to haveeliminated all signs of infection spontaneously. All of the infectedcontrol animals had gross evidence of pyelonephritis at autopsy. Mosthad renal and/or perinephric abscesses and intrarenal stone formation.Histologic examination demonstrated the presence of acute parenchymalinflammation with abscesses in all cases, except the one animal withsterile urine which had normal kidneys.

In contrast, the urine pH of the acetohydroxamic acid treated animalsremained in the normal range. Urine obtained at autopsy containedgreater than 10⁵ Proteus misabilis per ml but bladder stone formationwas minimal. Differences in mean urine pH and stone formation, betweenthe control and acetohydroxamic acid treated groups were satisticallysignificant (P.<0.01). Although all acetohydroxamic acid treated ratshad abundant Proteus mirabilis in their urine, none had gross evidenceof pyelonephritis or renal stone formation and nearly all had normalkidneys histologically. The results are summarized in Table 3.

                                      TABLE 3                                     __________________________________________________________________________    UNTREATED CONTROLS                                                            __________________________________________________________________________                    Urine  Autopsy                                                                              Stone                                           Animal                                                                            Urine                                                                              Urine Vol.                                                                           Acetohy. &                                                                           Urine  wt. Stone  Gross      Microscopic               No. pH   ml     mg/ml  bacteria/ml                                                                          mg  Composition                                                                          Kidney     Kidney                    __________________________________________________________________________    337 Expired                                                                   338 8.6   8            > 10.sup.5                                                                           25  Struvite                                                                             Bilat. Papillary                                                                         Pyelitis, Minimal                                                  Stones     Nephritis                 339 7.0  11            Sterile                                                                               1  Apatite                                                                              Normal     Normal                    340 8.1  17            > 10.sup.5                                                                           40  Struvite                                                                             Bilat. Papillary                                                                         Pyelonephritis                                                     Stones & Bilat.                                                               Pyelonephritis                       341 8.7  15            > 10.sup.5                                                                           41  Struvite                                                                             Bilat. Papillary                                                                         Pyelonephritis with                                                Stones & Bilat.                                                                          Papillary Necrosis                                                 Pyelonephritis                       342 8.7   7            > 10.sup.5                                                                           84  Struvite                                                                             Unilat. Pyelo-                                                                           Pyelonephritis with                                                nephritis with                                                                           Papillary Necrosis                                                 no stones                            343 8.6  11            > 10.sup.5                                                                           56  Struvite                                                                             Bilat. Pyelo-                                                                            Pyelonephritis                                                     nephritis with                                                                Bilat. Papillary                                                              Stones                               347 8.6  12            > 10.sup.5                                                                           25  Struvite                                                                             Unilat. Papillary                                                                        Pyelonephritis with                                                Stones, Contralat.                                                                       Papillary Necrosis                                                 Pyelonephritis                                                                with Abscess                         348 Expired                                                                   349 8.5  13            > 10.sup.5                                                                           94  Struvite                                                                             Bilat. Pyelo-                                                                            Pyelonephritis with                                                nephritis, Bilat.                                                                        Papillary Necrosis                                                 Papillary Stones                     __________________________________________________________________________    AHA TREATED                                                                   __________________________________________________________________________                    Urine  Autopsy                                                                              Stone                                           Animal                                                                            Urine                                                                              Urine Vol.                                                                           Acetohy. &                                                                           Urine  wt. Stone  Gross      Microscopic               No. pH   ml     mg/ml  bacteria/ml                                                                          mg  Composition                                                                          Kidney     Kidney                    __________________________________________________________________________    325 7.4  13      .3    > 10.sup.5                                                                           12  Struvite                                                                             Normal     Pyelonephritis            326 6.6  17     1.5    > 10.sup.5                                                                            4  Struvite                                                                             Normal     Mild Pyelitis, No                                                             Nephritis                 327 6.6  11     2.5    > 10.sup.5                                                                           21  Struvite                                                                             Normal     Normal                    328 6.7  12     1.3    > 10.sup.5                                                                            4  Struvite &                                                                           Normal     Normal                                                      Apatite                                     329 Expired                                                                   330 7.0   8      .9    > 10.sup.5                                                                           11  Struvite                                                                             Normal     Normal                    331 6.3  17     1.5    > 10.sup.5                                                                            1  Apatite                                                                              Normal     Normal                    332 6.5  20     1.6    > 10.sup.5                                                                           19  Struvite                                                                             Normal     Normal                    333 Expired                                                                   334 6.5   9     1.5    > 10.sup.5                                                                            1  Apatite                                                                              Normal     Mild Pyelitie                                                                 No Nephritis              335 6.3  16      .5    > 10.sup.5                                                                            3  Struvite                                                                             Normal     Normal                    336 6.4   8     2.5    > 10.sup.5                                                                            7  Apatite                                                                              Normal     Normal                    __________________________________________________________________________     pH, urine volume, and acetohydroxamic acid concentrations are average of      day individual values. All autopsy urine specimen contained Proteus           mirabilis, except animal No. 339.                                        

It can be seen from the data in Table 3, that where the infected ratswere given acetohydroxamic acid, while colonization of the urinary tractwith bacteria continued the tissue damage and virulence of the diseasedecreased because acetohydroxamic acid maintained in urine pH in thephysiological range. In addition it can be seen from the above data inTable 3 that the rats treated with the acetohydroxamic acid inaccordance with the present invention had significantly less bladderstone formation than the control rats. This therefore establishes theability of the acetohydroxamic acid not only to decrease stone formationand dissolve urinary tract stones but also to decrease or eliminate thepathogenicity of urinary colonization by urease producing bacteria.

While the present invention has been described primarily with regard tothe foregoing exemplification, it should be understood that the presentinvention cannot under any circumstances be deemed as limited theretobut rather must be construed as broadly as any and all equivalentsthereof.

What is claimed is:
 1. A method of treating urinary infections which comprises administering to a patient suffering from a urinary infection associated with presence in the urine of bacteria of the species Proteus, an effective anti-bacterial dosage of a composition comprising in combinationa. a dosage of methenamine which produces a concentration of methenamine in the patient's urine of from about 0.05 to about 1.0 mg/ml; and b. a dosage of acetohydroxamic acid which produces a concentration of hydroxamate groups in the patient's urine of from about 0.05 to about 2.0 mg/ml.
 2. The method of claim 1, wherein said methenamine is administered in a dosage of about 0.5 to about 8.0 grams/day.
 3. The method of claim 1, wherein said acetohydroxamic acid is administered in dosage of about 0.25 to about 4.0 grams/day.
 4. A composition adapted to be administered to a patient suffering from a bacterial urinary infection comprising in combination:a. a dosage of methenamine which produces a concentration of methenamine in the patient's urine of from about 0.05 to about 1.0 mg/ml; and b. a dosage of acetohydroxamic acid which produces a concentration of hydroxamate groups in the patient's urine of from about 0.05 to about 2.0 mg/ml.
 5. The composition of claim 4, wherein said methenamine is present in an amount to provide a dosage of about 0.5 to about 8.0 grams/day.
 6. The composition of claim 4, wherein said acetohydroxamic acid is present in an amount to provide a dosage of about 0.25 to about 4.0 grams/day. 